Extremely Low-Frequency Electromagnetic Fields Affect the miRNA-Mediated Regulation of Signaling Pathways in the GC-2 Cell Line.

PloS one · 2015 · Mixed · Low evidence

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MixedLow evidencePaper 4738

Study snapshot

PublicationPloS one · 2015

Study typein_vitro

Reported directionMixed

Evidence ratingLow

Abstract

Extremely low-frequency electromagnetic fields (ELF-EMFs) can affect male reproductive function, but the underlying mechanism of this effect remains unknown. miRNA-mediated regulation has been implicated as an important epigenetic mechanism for regulatory pathways. Herein, we profiled miRNA expression in response to ELF-EMFs in vitro. Mouse spermatocyte-derived GC-2 cells were intermittently exposed to a 50 Hz ELF-EMF for 72 h (5 min on/10 min off) at magnetic field intensities of 1 mT, 2 mT and 3 mT. Cell viability was assessed using the CCK-8 assay. Apoptosis and the cell cycle were analyzed with flow cytometry. miRNA expression was profiled using Affymetrix Mouse Genechip miRNA 3.0 arrays. Our data showed that the growth, apoptosis or cell cycle arrest of GC-2 cells exposed to the 50 Hz ELF-EMF did not significantly change. However, we identified a total of 55 miRNAs whose expression significantly changed compared with the sham group, including 19 differentially expressed miRNAs (7 miRNAs were upregulated, and 12 were downregulated) in the 1 mT exposure group and 36 (9 miRNAs were upregulated, and 27 were downregulated) in the 3 mT exposure group. The changes in the expression of 15 selected miRNAs measured by real-time PCR were consistent with the microarray results. A network analysis was used to predict core miRNAs and target genes, including miR-30e-5p, miR-210-5p, miR-196b-5p, miR-504-3p, miR-669c-5p and miR-455-3p. We found that these miRNAs were differentially expressed in response to different magnetic field intensities of ELF-EMFs. GO term and KEGG pathway annotation based on the miRNA expression profiling results showed that miRNAs may regulate circadian rhythms, cytokine-cytokine receptor interactions and the p53 signaling pathway. These results suggested that miRNAs could serve as potential biomarkers, and the miRNA-mediated regulation of signaling pathways might play significant roles in the biological effects of ELF-EMFs.

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AI evidence extraction

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Study typein_vitro

Effect directionmixed

PopulationMouse spermatocyte-derived GC-2 cell line

Sample sizeNot supplied

ExposureELF · 0.05 MHz · 72 h intermittent (5 min on/10 min off)

Evidence strengthLowConfidence: 78% · Peer reviewed: yes

Main findings

GC-2 cell growth/viability, apoptosis, and cell cycle arrest did not significantly change after intermittent 50 Hz ELF-EMF exposure at 1–3 mT for 72 h. In contrast, miRNA expression changed versus sham: 55 miRNAs were significantly altered overall, including 19 differentially expressed miRNAs at 1 mT (7 up, 12 down) and 36 at 3 mT (9 up, 27 down); RT-PCR of 15 selected miRNAs was consistent with microarray results. Bioinformatic analyses suggested potential involvement of circadian rhythm, cytokine-cytokine receptor interaction, and p53 signaling pathways.

Outcomes measured

Cell viability (CCK-8)
Apoptosis (flow cytometry)
Cell cycle (flow cytometry)
miRNA expression profiling (Affymetrix Mouse Genechip miRNA 3.0 arrays; RT-PCR validation)
Predicted signaling pathways/functional annotation (GO, KEGG; network analysis)

Limitations

In vitro cell-line study; findings may not generalize to in vivo male reproductive function
No sample size/replication details provided in abstract
Pathway and target-gene implications are based on prediction/annotation rather than direct functional validation
2 mT exposure group mentioned in methods but differential miRNA results for 2 mT not reported in abstract

View raw extracted JSON

{
  "study_type": "in_vitro",
  "exposure": {
    "band": "ELF",
    "source": null,
    "frequency_mhz": 0.05,
    "sar_wkg": null,
    "duration": "72 h intermittent (5 min on/10 min off)"
  },
  "population": "Mouse spermatocyte-derived GC-2 cell line",
  "sample_size": null,
  "outcomes": [
    "Cell viability (CCK-8)",
    "Apoptosis (flow cytometry)",
    "Cell cycle (flow cytometry)",
    "miRNA expression profiling (Affymetrix Mouse Genechip miRNA 3.0 arrays; RT-PCR validation)",
    "Predicted signaling pathways/functional annotation (GO, KEGG; network analysis)"
  ],
  "main_findings": "GC-2 cell growth/viability, apoptosis, and cell cycle arrest did not significantly change after intermittent 50 Hz ELF-EMF exposure at 1–3 mT for 72 h. In contrast, miRNA expression changed versus sham: 55 miRNAs were significantly altered overall, including 19 differentially expressed miRNAs at 1 mT (7 up, 12 down) and 36 at 3 mT (9 up, 27 down); RT-PCR of 15 selected miRNAs was consistent with microarray results. Bioinformatic analyses suggested potential involvement of circadian rhythm, cytokine-cytokine receptor interaction, and p53 signaling pathways.",
  "effect_direction": "mixed",
  "limitations": [
    "In vitro cell-line study; findings may not generalize to in vivo male reproductive function",
    "No sample size/replication details provided in abstract",
    "Pathway and target-gene implications are based on prediction/annotation rather than direct functional validation",
    "2 mT exposure group mentioned in methods but differential miRNA results for 2 mT not reported in abstract"
  ],
  "evidence_strength": "low",
  "confidence": 0.78,
  "peer_reviewed_likely": "yes",
  "keywords": [
    "ELF-EMF",
    "50 Hz",
    "magnetic field",
    "mT",
    "GC-2",
    "spermatocyte",
    "male reproductive function",
    "miRNA",
    "microarray",
    "RT-PCR",
    "p53 signaling",
    "circadian rhythm",
    "cytokine-cytokine receptor interaction"
  ],
  "suggested_hubs": [
    {
      "slug": "occupational-exposure",
      "weight": 0.25,
      "reason": "Study uses 50 Hz ELF magnetic fields at mT levels, relevant to some occupational ELF exposure contexts (though source not specified)."
    }
  ]
}

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AI-extracted fields are generated from the abstract and metadata and may be incomplete or incorrect. This material is for informational purposes and is not medical advice.

Identifiers and source

DOI: 10.1371/journal.pone.0139949
PubMed: 26439850

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